Double-stranded RNA compounds to CASP2 and uses thereof

ABSTRACT

The present disclosure relates to methods of treating a patient suffering from or at risk of developing an ocular disease, disorder or injury, and includes treatment regimens using a double-stranded RNA compound that down-regulates CASP2 expression, or a pharmaceutically acceptable salt thereof.

RELATED APPLICATIONS

This application is a continuation application of and claims priority toU.S. application Ser. No. 14/370,061, filed on Jul. 1, 2014, now U.S.Pat. No. 9,382,542, which is the U.S. National Stage of InternationalApplication No. PCT/US2013/020012, filed on Jan. 3, 2013, which waspublished in English under PCT Article 21(2), which claims the benefitof U.S. Provisional Application Ser. No. 61/582,886 filed Jan. 4, 2012and of U.S. Provisional Application Ser. No. 61/596,231 filed Feb. 8,2012, both entitled “Methods for Treating Eye Disorders”, each of theseapplications is incorporated herein by reference in its entirety and forall purposes.

SEQUENCE LISTING

This application incorporates-by-reference nucleotide and/or amino acidsequences which are present in the file named “243-PCT1.ST25.txt”, whichis 25 kilobytes in size, and which was created Jan. 3, 2013 in theIBM-PCT machine format, having an operating system compatibility withMS-Windows.

FIELD OF THE INVENTION

Provided herein are compositions and methods of treating ocular disease,ocular disorder or ocular injury.

BACKGROUND OF THE INVENTION

There is a general lack of therapies for optic neuropathies. Glaucoma istreated in part by lowering intraocular pressure. Optic neuritis ismanaged by corticosteroids, but this does not affect the long-termcourse of the disease. Compressive optic neuropathy is treated byremoving the tumor or aneurysm pressing on the optic nerve or chiasm.All other optic neuropathies, including nonarteritic anterior ischemicoptic neuropathy (NAION), represent unmet medical needs. (Levin La.Axonal loss and neuroprotection in optic neuropathies. Can J Ophthalmol.2007, 42(3):403-8).

PCT Publication Nos. WO 2008/050329 and WO 2009/044392 are directed toinhibitors of pro-apoptotic genes and disclose double-stranded RNAmolecules targeting, inter alia, Caspase 2.

PCT Publication No. WO 2010/048352 is directed to compositions andmethods of treating ocular diseases and discloses, inter alia, thechemically modified, double-stranded RNA compound QPI-1007 targeting theCaspase 2 gene.

Ahmed Z. et al., (Cell Death and Disease (2011) 2, e173) suggest thatretinal ganglion cell (RGC) apoptosis induced by optic nerve injury in arat model of optic nerve transection, involves activation of Caspase 2,and that synthetic double stranded RNA compounds designed to inhibitexpression of Caspase 2 represent potential neuroprotective agents forintervention in human diseases involving RGC loss.

SUMMARY OF THE INVENTION

Provided herein are compounds and compositions for use in treating asubject and methods of treating a subject, wherein the subject issuffering from or at risk of developing an ocular disease, oculardisorder or ocular injury. The treatment comprises administering to thesubject's eye a therapeutically effective dose of a double-stranded RNAcompound that down regulates Caspase 2 (CASP2) expression in a singletreatment or according to a treatment regimen having a dosing intervalselected from one week, two weeks, one month, six weeks, two months andlonger, or combinations thereof, wherein the regimen is maintained untilthe desired therapeutic effect is achieved for the subject. The dosesprovided herein present a favorable outcome to patients suffering fromor at risk of developing an ocular disease, ocular disorder or ocularinjury or in need of ocular neuroprotection, such as for examplepatients suffering from or at risk of developing non-arteritic anteriorischemic optic neuropathy (NAION) and other optic neuropathies, such asglaucoma, that result in the death of retinal ganglion cells (RGCs). Thetreatment regimens provided herein present a favorable outcome topatients suffering from or at risk of developing an ocular disease,ocular disorder or ocular injury including, and without being limitedto, NAION and glaucoma.

Various aspects and embodiments provided herein involve use of a nucleicacid molecule, or a pharmaceutically acceptable salt thereof, thatdown-regulates expression of CASP2 in a subject's eye, that bind anucleotide sequence (such as an mRNA sequence) or portion thereof,encoding CASP2, for example, the mRNA coding sequence (SEQ ID NO:3-5)for human CASP2, encoding one or more proteins or protein subunitsexemplified by SEQ ID NO:6-8. In some embodiments, a nucleic acidmolecule or a pharmaceutically acceptable salt thereof, that downregulates, or targets expression of the CASP2 gene is administered to aneye of the subject at a dose of about 0.05 mg to about 10 mg per eye,such as about 0.2 mg to about 6.0 mg per eye. In various embodiments thenucleic acid molecule or a pharmaceutically acceptable salt thereof isadministered as an intravitrcal (IVT) injection. In some embodiments thenucleic acid molecule or a pharmaceutically acceptable salt thereof isadministered unilaterally. In some embodiments the nucleic acid moleculeor a pharmaceutically acceptable salt thereof is administeredbilaterally. In preferred embodiments the nucleic acid molecule is adouble-stranded RNA (dsRNA) compound comprising an antisense strand anda sense strand. In various embodiments, the double-stranded RNA compoundcomprises an antisense strand with the sequence: 5′ AGGAGUUCCACAUUCUGGC3′ (SEQ ID NO: 2) and a sense strand with the sequence 5′GCCAGAAUGUGGAACUCCU 3′ (SEQ ID NO: 1).

In preferred embodiments, the double-stranded RNA compound (referred toherein as “QPI-1007”) has the following structure:

(sense strand; SEQ ID NO: 1) 5′ iB - GCCAGAAUGUGGAACUCCU 3′(antisense strand; SEQ ID NO: 2) 3′ CGGUCUUACACCUUGAGGA 5′wherein each A, C, U and G is a nucleotide and each consecutivenucleotide is joined to the next nucleotide by a phosphodiester bond;wherein each nucleotide is independently an unmodified ribonucleotide, a2′-O-Methyl sugar modified ribonucleotide or a L-DNA nucleotide;wherein the sense strand comprises, counting from the 5′ terminus, anunmodified ribonucleotide at each of positions 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17 and 19 and a L-deoxycytidine atposition 18, and an inverted abasic deoxyribose cap (iB) covalentlybound to the 5′ terminus; andwherein the antisense strand comprises, counting from the 5′ terminus, a2′-O-Methyl sugar modified ribonucleotide at each of positions 2, 4, 6,8, 11, 13, 15, 17 and 19 and an unmodified ribonucleotide at each ofpositions 1, 3, 5, 7, 9, 10, 12, 14, 16 and 18.

In preferred embodiments, there is provided a double-stranded RNAcompound, or a pharmaceutically acceptable salt thereof, having thestructure:

(sense strand; SEQ ID NO: 1) 5′ iB - GCCAGAAUGUGGAACUCCU 3′(antisense strand; SEQ ID NO: 2) 3′ CGGUCUUACACCUUGAGGA 5′wherein each A, C, U and G is a nucleotide and each consecutivenucleotide is joined to the next nucleotide by a phosphodiester bond;wherein each nucleotide is independently an unmodified ribonucleotide, a2′-O-Methyl sugar modified ribonucleotide or a L-DNA nucleotide;wherein the sense strand comprises, counting from the 5′ terminus, anunmodified ribonucleotide at each of positions 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17 and 19 and a L-deoxycytidine atposition 18, and an inverted abasic deoxyribose cap (iB) covalentlybound to the 5′ terminus;wherein the antisense strand comprises, counting from the 5′ terminus, a2′-O-Methyl sugar modified ribonucleotide at each of positions 2, 4, 6,8, 11, 13, 15, 17 and 19 and an unmodified ribonucleotide at each ofpositions 1, 3, 5, 7, 9, 10, 12, 14, 16 and 18;for use in the treatment of a patient suffering from or at risk ofdeveloping an ocular disease, an ocular disorder or an ocular injury;wherein the compound, or salt thereof, is administered to the patient'seye at a dose of about 0.05 mg to about 10 mg per eye.

In various embodiments, QPI-1007 is to be administered to a patient'seye at a dose of about 0.05 mg to about 10 mg per eye, at a dose ofabout 0.2 mg to about 6.0 mg per eye or at a dose of about 2.4 mg toabout 6.0 mg per eye.

In various embodiments, the double-stranded RNA compound is administeredto the patient's eye at a dose of about 0.05 mg, 0.06 mg, 0.07 mg, 0.08mg, 0.09 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8mg, 0.9 mg, 1.0 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7mg, 1.8 mg, 1.9 mg, 2.0 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6mg, 2.7 mg, 2.8 mg, 2.9 mg, 3.0 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4.0 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5.0 mg, 5.1 mg, 5.2 mg, 5.3mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6.0 mg, 6.1 mg, 6.2mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7.0 mg, 7.1mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8.0mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9mg, 9.0 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8mg, 9.9 mg, or about 10.0 mg, or at a dose of 0.05 mg, 0.06 mg, 0.07 mg,0.08 mg, 0.09 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7mg, 0.8 mg, 0.9 mg, 1.0 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6mg, 1.7 mg, 1.8 mg, 1.9 mg, 2.0 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3.0 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4.0 mg, 4.1 mg, 4.2 mg, 4.3mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5.0 mg, 5.1 mg, 5.2mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6.0 mg, 6.1mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7.0mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9mg, 8.0 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8mg, 8.9 mg, 9.0 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7mg, 9.8 mg, 9.9 mg or 10.0 mg.

In certain embodiments of the compound for use or the method oftreating, the double-stranded RNA compound is administered at a dose ofabout 0.2 mg per eye. In some embodiments of the compound for use or themethod of treating, the double-stranded RNA compound is administered ata dose of 0.2 mg per eye.

In certain embodiments of the compound for use or the method oftreating, the double-stranded RNA compound is administered at a dose ofabout 0.6 mg per eye. In some embodiments of the compound for use or themethod of treating, the double-stranded RNA compound is administered ata dose of 0.6 mg per eye.

In certain embodiments of the compound for use or the method oftreating, the double-stranded RNA compound is administered at a dose ofabout 1.2 mg per eye. In some embodiments of the compound for use or themethod of treating, the double-stranded RNA compound is administered ata dose of 1.2 mg per eye.

In certain embodiments of the compound for use or the method oftreating, the double-stranded RNA compound is administered at a dose ofabout 2.4 mg per eye. In some embodiments of the compound for use or themethod of treating, the double-stranded RNA compound is administered ata dose of 2.4 mg per eye.

In certain embodiments of the compound for use or the method oftreating, the double-stranded RNA compound is administered at a dose ofabout 4.8 mg per eye. In some embodiments of the compound for use or themethod of treating, the double-stranded RNA compound is administered ata dose of 4.8 mg per eye.

In certain embodiments of the compound for use or the method oftreating, the double-stranded RNA compound is administered at a dose ofabout 6.0 mg per eye. In some embodiments of the compound for use or themethod of treating, the double-stranded RNA compound is administered ata dose of 6.0 mg per eye.

In preferred embodiments of the compound for use or the method oftreating, the compound, or a pharmaceutically acceptable salt thereof,is prepared for intravitreal (IVT) injection. In preferred embodimentsof the compound for use or the method of treating, the compound, or apharmaceutically acceptable salt thereof, is administered as anintravitreal (IVT) injection. In some embodiments, the intravitreal(IVT) injection is administered in a single treatment. In someembodiments of the compound for use or the method of treating, thetreatment comprises multiple (i.e. 2, 3, 4, 5, 6 or more)administrations of the double-stranded RNA compound. In preferredembodiments, the treatment comprises a multiple dose regimen, forexample multiple (i.e. 2, 3, 4, 5, 6 or more) consecutiveadministrations. In some embodiments, the multiple administrationscomprise multiple intravitreal (IVT) injections. In some embodiments ofthe compound for use or the method of treating provided herein, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is administered as six (6) consecutive intravitreal (IVT)injections. As provided herein, the multiple administrations can occurat regular intervals or at irregular intervals. In preferredembodiments, the multiple administrations occur at regular intervals. Asprovided herein, the regular intervals are selected from the groupconsisting of about one week, two weeks, one month, six weeks, twomonths and longer than two months. In some embodiments, the regularintervals are of about one month. In preferred embodiments of thecompound for use or the method of treating, the double-stranded RNAcompound is administered as intravitreal (IVT) injections at regularintervals of one month for six (6) consecutive months.

In some embodiments of the compound for use or the method of treating,for example in the treatment of a chronic eye disease, thedouble-stranded RNA compound is administered as intravitreal (IVT)injections at regular intervals of one month or two months for more thansix (6) months, for example up to 12 months, or 24 months or more.

In certain embodiments of the compound for use or the method oftreating, the volume of a single intravitreal (IVT) injection is about 1μl to about 200 μl (μl refers to microliter). In some embodiments, theinjection volume is about 5 μl to about 200 μl. In some embodiments, theinjection volume is about 20 μl to about 200 μl. In some embodiments,the injection volume is between about 50 μl to about 100 μl. In someembodiments, the volume of a single intravitreal (IVT) injection is 50μl or 100 μl.

In various embodiments of the compound for use or the method oftreating, the double-stranded RNA compound, or a pharmaceuticallyacceptable salt thereof, is useful in the treatment of a subjectsuffering from or at risk of developing an ocular disease, an oculardisorder or an ocular injury, including for example, visual field loss,visual acuity loss, neurodegeneration, increased intraocular pressure,an ischemic event, retinal injury or optic nerve injury. In someembodiments of the compound for use or the method of treating, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is useful in the treatment of a subject suffering from or atrisk of developing retinal injury or optic nerve injury. In someembodiments provided herein, the retinal injury or optic nerve injurycomprises ischemic or hypoxic injury. In some embodiments of thecompound for use or the method of treating, the double-stranded RNAcompound, or a pharmaceutically acceptable salt thereof, is useful inachieving neuroprotection in the eye of the subject, for exampleneuroprotection of the RGC and/or neuroprotection of the optic nerve. Insome embodiments of the compound for use or the method of treating, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is useful in reducing or preventing visual field loss in theeye of the subject. In some embodiments of the compound for use or themethod of treating, the double-stranded RNA compound, or apharmaceutically acceptable salt thereof, is useful in reducing orpreventing visual acuity loss in the eye of the subject.

In some embodiments of the compound for use or the method of treating,the double-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is useful in increasing or enhancing visual acuity in the eyeof the subject, in particular in the eye of a NAION or glaucoma patient.

In various embodiments of the compound for use or the method oftreating, the ocular disease, ocular disorder, or ocular injury isselected from the group consisting of ocular neuropathy, elevatedintraocular pressure (IOP), glaucoma, acute angle closure (AAC), acuteangle closure glaucoma (AACG), primary angle closure disease (PACD),primary angle closure glaucoma (PACG), dry eye, Sjögrens Syndrome,diabetic retinopathy (DR), diabetic macular edema (DME), age relatedmacular degeneration (AMD), optic neuritis, central retinal veinocclusion, brunch retinal vein occlusion, ischemic optic neuropathy,optic nerve atrophy, optic nerve injury, non-arteritic anterior ischemicoptic neuropathy (NAION), retinopathy of prematurity (ROP), retinitispigmentosa (RP), retinal degeneration, retinal ganglion degeneration,macular degeneration, hereditary optic neuropathy, Leber's hereditaryoptic neuropathy, metabolic optic neuropathy, neuropathy due to a toxicagent, all secondary glaucomas, ocular hypertension, normal tensionglaucoma, and a neuropathy caused by an adverse drug reaction or avitamin deficiency.

In certain embodiments of the compound for use or the method oftreating, the ocular disease, ocular disorder or ocular injury is opticnerve atrophy. In some embodiments, the optic nerve atrophy is chronicoptic nerve atrophy.

In certain embodiments of the compound for use or the method oftreating, the ocular disease, ocular disorder or ocular injury isretinal degeneration.

In certain embodiments of the compound for use or the method oftreating, the ocular disease, ocular disorder or ocular injury isnon-arteritic anterior ischemic optic neuropathy (NAION). In someembodiments, the NAION is acute NAION. In some embodiments of thecompound for use or the method of treating, the ocular disease, oculardisorder or ocular injury is NAION or acute NAION and thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is administered to the patient's eye within 14 days of theonset of NAION symptoms. In some embodiments of the compound for use orthe method of treating, the ocular disease, ocular disorder or ocularinjury is NAION or acute NAION and the double-stranded RNA compound, ora pharmaceutically acceptable salt thereof, is administered to thepatient's eye within 28 days of the onset of NAION symptoms. In certainembodiments of the compound for use or the method of treating, theocular disease, ocular disorder or ocular injury is optic neuritis.

In some embodiments of the compound for use or the method of treating,the ocular disease, ocular disorder or ocular injury is glaucoma, forexample, a primary glaucoma or a secondary glaucoma. In someembodiments, the glaucoma is a primary glaucoma selected from the groupconsisting of primary open angle glaucoma, normal-tension glaucoma,primary angle-closure glaucoma (PACG), acute angle-closure glaucoma(AACG) and angle-closure glaucoma. In some embodiments of the compoundfor use or the method, the ocular disease, ocular disorder or ocularinjury is primary angle closure (PAC) or acute angle closure (AAC). Insome embodiments, the glaucoma is secondary glaucoma selected from thegroup consisting of pseudoexfoliation glaucoma, pigmentary glaucoma,neovascular glaucoma, steroid-induced glaucoma, and treatment refractoryglaucoma.

In certain embodiments of the compound for use or the method oftreating, the ocular disease, ocular disorder or ocular injury isLeber's hereditary optic neuropathy.

In various embodiments of the compound for use or the method of treatingprovided herein, the double-stranded RNA compound is in a form of apharmaceutically acceptable salt. In preferred embodiments, thepharmaceutically acceptable salt is a sodium salt. A sodium salt of thecompound means any compound containing at least one sodium atom.

In various embodiments of the compound for use or the method of treatingprovided herein, the double-stranded RNA compound is present in acomposition that comprises the compound and a pharmaceuticallyacceptable carrier. In some embodiments, the pharmaceutically acceptablecarrier is sterile saline solution suitable for injection into the eye.In certain embodiments, the composition further comprises apreservative. In various embodiments, the composition is formulated as acream, a foam, a paste, an ointment, an emulsion, a liquid solution, aneye drop, a gel, spray, a suspension, a microemulsion, microspheres,microcapsules, nanospheres, nanoparticles, lipid vesicles, liposomes,polymeric vesicles, a patch, or a contact lens. In preferredembodiments, the composition is formulated as a liquid solution. In someembodiments, the liquid solution is prepared for a single doseintravitreal (IVT) injection and the volume of a single dose IVTinjection is between about 20 μl to about 200 μl, preferably 50 μl toabout 100 μl. In some embodiments, the liquid solution is prepared for asingle dose intravitreal (IVT) injection and the volume of a single doseIVT injection is 100 μl. In some embodiments, the liquid solution isprepared for a single dose intravitreal (IVT) injection and the volumeof a single dose IVT injection is 50 μl.

In another aspect, provided herein is an injectable compositioncomprising a pharmacologically acceptable aqueous excipient and thedouble-stranded RNA compound described hereinabove and infra. In variousembodiments provided herein, the injectable composition is for use intreatment of a patient suffering from or at risk for developing anocular disease, an ocular disorder or an ocular injury and foradministering to the patient's eye.

In another aspect provided herein is a sodium salt of a double-strandedRNA compound targeting Caspase 2 having the structure:

(sense strand; SEQ ID NO: 1) 5′ iB - GCCAGAAUGUGGAACUCCU 3′(antisense strand; SEQ ID NO: 2) 3′ CGGUCUUACACCUUGAGGA 5′wherein each A, C, U and G is a nucleotide and each consecutivenucleotide is joined to the next nucleotide by a phosphodiester bond;wherein the sense strand comprises, counting from the 5′ terminus, anunmodified ribonucleotide at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17 and 19, a L-deoxycytidine at position 18, andan inverted abasic deoxyribose moiety 5′ cap; andwherein the antisense strand comprises, counting from the 5′ terminus, a2′-O-Methyl sugar modified ribonucleotide at positions 2, 4, 6, 8, 11,13, 15, 17 and 19 and an unmodified ribonucleotide at positions 1, 3, 5,7, 9, 10, 12, 14, 16 and 18;wherein the molecular formula is C₃₇₅ H₄₃₉ N₁₄₃ Na₃₇ O₂₆₆ P₃₇; andwherein the molecular weight is 13,202 Da.

In another aspect, provided herein is a pharmaceutical compositioncomprising a sodium salt of double-stranded RNA compound having thestructure:

(sense strand; SEQ ID NO: 1) 5′ iB - GCCAGAAUGUGGAACUCCU 3′(antisense strand; SEQ ID NO: 2) 3′ CGGUCUUACACCUUGAGGA 5′wherein each A, C, U and G is a nucleotide and each consecutivenucleotide is joined to the next nucleotide by a phosphodiester bond;wherein the sense strand comprises, counting from the 5′ terminus, anunmodified ribonucleotide at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17 and 19, a L-deoxycytidine at position 18, andan inverted abasic deoxyribose moiety 5′ cap; andwherein the antisense strand comprises, counting from the 5′ terminus, a2′-O-Methyl sugar modified ribonucleotide at positions 2, 4, 6, 8, 11,13, 15, 17 and 19 and an unmodified ribonucleotide at positions 1, 3, 5,7, 9, 10, 12, 14, 16 and 18;wherein the molecular formula is C₃₇₅ H₄₃₉ N₁₄₃ Na₃₇ O₂₆₆ P₃₇ and themolecular weight is 13,202 Da; anda pharmaceutically acceptable excipient or carrier or mixture thereof.

In various embodiments, the sodium salt of the double-stranded RNAcompound as provided herein is present in the composition at an amountof about 0.05 mg to about 10.0 mg per dosage form. In some embodiments,the sodium salt of the double-stranded RNA compound as provided hereinis present in the composition in an amount of 0.05 mg to 10.0 mg perdosage unit.

In certain embodiments of the pharmaceutical composition, the sodiumsalt of QPI-1007 is present in the composition in an amount of about 0.1mg to about 8.0 mg per dosage form, or at in an amount of 0.1 mg to 8.0mg. In certain embodiments, the sodium salt of QPI-1007 is present inthe composition in an amount of about 0.2 mg to about 6.0 mg per dosageform, or at in an amount of 0.2 mg to 6.0 mg.

In certain embodiments of the pharmaceutical composition, the sodiumsalt of QPI-1007 is present in the composition in an amount of about0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1.0 mg, 1.1 mg, 1.2 mg, 1.3mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2.0 mg, 2.1 mg, 2.2mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3.0 mg, 3.1mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4.0mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9mg, 5.0 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8mg, 5.9 mg, 6.0 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7mg, 6.8 mg, 6.9 mg, 7.0 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6mg, 7.7 mg, 7.8 mg, 7.9 mg, 8.0 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9.0 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, or 10.0 mg per dose form, orin an amount of 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.1 mg, 0.2mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1.0 mg, 1.1mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2.0mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9mg, 3.0 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8mg, 3.9 mg, 4.0 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7mg, 4.8 mg, 4.9 mg, 5.0 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6mg, 5.7 mg, 5.8 mg, 5.9 mg, 6.0 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7.0 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8.0 mg, 8.1 mg, 8.2 mg, 8.3mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9.0 mg, 9.1 mg, 9.2mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, or 10.0 mgper dose form.

In certain embodiments of the pharmaceutical composition, the sodiumsalt of QPI-1007 is present in the composition in an amount of about 0.2mg per dose form, or in an amount of 0.2 mg per dose form.

In certain embodiments of the pharmaceutical composition, the sodiumsalt of QPI-1007 is present in the composition in an amount of about 0.6mg per dose form, or in an amount of 0.6 mg per dose form.

In certain embodiments of the pharmaceutical composition, the sodiumsalt of QPI-1007 is present in the composition in an amount of about 1.2mg per dose form, or in an amount of 1.2 mg per dose form.

In certain embodiments of the pharmaceutical composition, the sodiumsalt of QPI-1007 is present in the composition in an amount of about 2.4mg per dose form, or in an amount of 2.4 mg per dose form.

In certain embodiments, the sodium salt of QPI-1007 is present in thecomposition in an amount of about 4.8 mg per dose form, or in an amountof 4.8 mg per dose form.

In certain embodiments of the pharmaceutical composition, the sodiumsalt of QPI-1007 is present in the composition in an amount of about 6.0mg per dose form, or in an amount of 6.0 mg per dose form.

In certain embodiments of the pharmaceutical composition, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is formulated in a pharmaceutically acceptable excipient orcarrier at a concentration of about 0.5 mg/mL to about 100.0 mg/mL, orat a concentration of 0.5 mg/mL to 100.0 mg/mL. In certain embodimentsof the pharmaceutical composition, the double-stranded RNA compound, ora pharmaceutically acceptable salt thereof, is formulated in apharmaceutically acceptable excipient or carrier at a concentration ofabout 1.0 mg/mL to about 80.0 mg/mL, or at a concentration of 1.0 mg/mLto 80.0 mg/mL In certain embodiments, the double-stranded RNA compound,or a pharmaceutically acceptable salt thereof, is formulated in apharmaceutically acceptable excipient or carrier at a concentration ofabout 2.0 mg/mL to about 60.0 mg/mL, or at a concentration of 2.0 mg/mLto 60.0 mg/mL.

In certain embodiments of the pharmaceutical composition, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is present in the composition at a concentration of about 2.0mg/mL, or at a concentration of 2.0 mg/mL. In certain embodiments, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is present in the composition at a concentration of about 12.0mg/mL, or at a concentration of 12.0 mg/mL. In certain embodiments, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is present in the composition at a concentration of about 24.0mg/mL, or at a concentration of 24.0 mg/mL. In certain embodiments, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is present in the composition at a concentration of about 48.0mg/mL, or at a concentration of 48.0 mg/mL. In certain embodiments, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is present in the composition at a concentration of about 60.0mg/mL, or at a concentration of 60.0 mg/mL In certain embodiments, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is present in the composition at a concentration of about 100.0mg/mL, or at a concentration of 100.0 mg/mL In certain embodiments, thedouble-stranded RNA compound, or a pharmaceutically acceptable saltthereof, is present in the composition at a concentration of about 120.0mg/mL, or at a concentration of 120.0 mg/mL.

In certain embodiments of the pharmaceutical composition, QPI-1007 or asodium salt thereof, is formulated in a pharmaceutically acceptableexcipient or carrier or mixture thereof at a concentration of about 60mg/mL. In certain embodiments, QPI-1007, or a pharmaceuticallyacceptable salt thereof, is formulated in a preservative-free, sterilesolution at a concentration of 60 mg/mL In certain embodiments thesterile solution is a phosphate-buffered saline.

In some embodiments, the compound for use described herein is present ina kit comprising the double-stranded RNA compound, or a pharmaceuticallyacceptable salt thereof, or the composition described hereinabove andinfra, and instructions for use thereof. In various embodiments of thekit, the use is for treatment of an ocular disease, an ocular disorderor an ocular injury, for example, wherein the ocular injury includesischemic injury, ischemia-reperfusion injury, mechanical injury, injuryor interruption of nerve fibers and/or is associated with lack of supplyof neurotrophic factor, or wherein the ocular disease, an oculardisorder or an ocular injury is associated with death of retinalganglion cells (RGCs). In various embodiments of the kit, the disease isselected from the group of diseases and disorders described herein. Invarious embodiments of the kit, the kit contains dosage units ofmedication of the double-stranded RNA compound, or a pharmaceuticallyacceptable salt thereof, or the composition described herein.

In various embodiments, the kit for use in the treatment of an oculardisease, an ocular disorder or an ocular injury comprises thedouble-stranded RNA compound or a pharmaceutically acceptable saltthereof, or the composition described hereinabove and infra, packaged ina suitable sealed container; at least one syringe needle, suitable forintravitreal injection; and at least one syringe. In some embodiments ofthe kit, the syringe is a precisely calibrated syringe. In someembodiments of the kit, the needle is a self-sealing syringe needle,suitable for intravitreal injection. In some embodiments of the kit, thekit for use in the treatment of an ocular disease, an ocular disorder oran ocular injury comprises the double-stranded RNA compound or apharmaceutically acceptable salt thereof, or the composition describedhereinabove and infra, packaged in a suitable scaled container; at leastone self-scaling syringe needle, suitable for intravitreal injection;and at least one precisely calibrated syringe. In various embodiments ofthe kit, the needle is selected from a 30-gauge, a 31-gauge, and a32-gauge needle. In various embodiments, the kit further comprisesprinted informational material describing the double-stranded RNAcompound or a pharmaceutically acceptable salt thereof, or thecomposition, its method of administration and any required safety andefficacy information as may be required by government regulations.

In a related aspect, provided are compositions or kits that include theQPI-1007 compound or a pharmaceutically acceptable salt thereof,packaged for use by a patient. The package may be labeled or include apackage label or insert that indicates the content of the package andprovides certain information regarding how the compound should be or canbe used by a patient, for example the label may include dosinginformation and/or indications for use. In certain embodiments thecontents of the label will bear a notice in a form prescribed by agovernment agency, for example the United States Food and Drugadministration. In certain embodiments, the label may indicate that thecompound is suitable for use in treating a patient suffering from adisease associated with increased expression of Caspase 2 and/orapoptosis of a retinal ganglion cell (RGC) and/or optic nerve damage;for example, the label may indicate that the compound is suitable foruse in treating NAION or glaucoma; or for example the label may indicatethat the compound or a pharmaceutically acceptable salt thereof, issuitable for use in treating an eye disease selected from the groupconsisting of ocular neuropathy, elevated intraocular pressure (IOP),glaucoma, acute angle closure (AAC), acute angle closure glaucoma(AACG), primary angle closure (PAC), primary angle closure glaucoma(PACG), dry eye, Sjögrens Syndrome, diabetic retinopathy (DR), diabeticmacular edema (DME), age related macular degeneration (AMD), opticneuritis, central retinal vein occlusion, brunch retinal vein occlusion,ischemic optic neuropathy, optic nerve atrophy, optic nerve injury,non-arteritic anterior ischemic optic neuropathy (NAION), retinopathy ofprematurity (ROP), retinitis pigmentosa (RP), retinal degeneration,retinal ganglion degeneration, macular degeneration, hereditary opticneuropathy, Leber's hereditary optic neuropathy, metabolic opticneuropathy, neuropathy due to a toxic agent, all secondary glaucomas,ocular hypertension, normal tension glaucoma, and a neuropathy caused byan adverse drug reaction or a vitamin deficiency.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is bar graph showing the proportion of NAION subjects who gainedor lost ≥3 lines of VA after receiving a single IVT injection ofQPI-1007 by cohort compared with historical controls at months 3 and 6.

FIG. 2 provides a flow chart outlining the Dose Escalation program

DETAILED DESCRIPTION OF THE INVENTION Definitions

For convenience certain terms employed in the specification, examplesand claims are described herein.

It is to be noted that, as used herein, the singular forms “a”, “an” and“the” include plural forms unless the content clearly dictatesotherwise.

As used herein the term “about” with regard to a numerical value refersto the numerical value ±10%.

As used herein, the term “inhibit”, “down-regulate”, or “reduce” withrespect to gene expression means the expression of the gene, or level ofRNA molecules or equivalent RNA molecules encoding one or more proteinsor protein subunits (e.g., mRNA), or activity of one or more proteins orprotein subunits, is reduced below that observed in the absence of aninhibitory factor (such as a nucleic acid molecule, e.g., an siNA, forexample having structural features as described herein); for example theexpression may be reduced to 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%,10%, 5% or less than that observed in the absence of an inhibitor.

“Nucleotide” is meant to encompass deoxyribonucleotides andribonucleotides, which may be natural or synthetic and modified orunmodified. Nucleotides include known nucleotide analogues, which aresynthetic, naturally occurring, and non-naturally occurring. Examples ofsuch analogs include, without limitation, phosphorothioates,phosphoramidites, methyl phosphonates, chiral-methyl phosphonates,2′-O-methyl ribonucleotides, and peptide-nucleic acids (PNAs).Modifications include changes to the sugar moiety, the base moiety andor the linkages between ribonucleotides in the oligoribonucleotide. Asused herein, the term “ribonucleotide” encompasses natural andsynthetic, unmodified and modified ribonucleotides and ribonucleotideanalogs which are synthetic, naturally occurring, and non-naturallyoccurring. Modifications include changes to the sugar moiety, to thebase moiety and/or to the linkages between ribonucleotides in theoligonucleotide.

A “mirror nucleotide” is a nucleotide with reversed chirality to thenaturally occurring or commonly employed nucleotide, i.e., a mirrorimage (L-nucleotide) of the naturally occurring D-nucleotide, referredto as L-RNA for a mirror ribonucleotide. The L-ribonucleotide orL-deoxyribonucleotide may further comprise at least one sugar, base andor backbone modification. See U.S. Pat. No. 6,586,238. U.S. Pat. No.6,602,858 discloses nucleic acid catalysts comprising at least oneL-nucleotide substitution. Mirror nucleotide includes for example L-DNA(L-deoxyriboadenosine-3′-phosphate (mirror dA);L-deoxyribocytidine-3′-phosphate (mirror dC);L-deoxyriboguanosine-3′-phosphate (mirror dG);L-deoxyribothymidine-3′-phosphate (mirror dT) and L-RNA(L-riboadenosine-3′-phosphate (mirror rA); L-ribocytidine-3′-phosphate(mirror rC); L-riboguanosine-3′-phosphate (mirror rG);L-ribouridine-3′-phosphate (mirror dU).

Ocular Diseases

In various embodiments the double-stranded RNA compounds provided hereinare useful in treating patients suffering from ocular diseases,disorders and injury in which neuroprotection of the optic nerve wouldbe of benefit, for example, without being limited to, in treating ION,including NAION, glaucoma, including glaucomatous optic neuropathy,Leber's hereditary optic neuropathy (LHON), or Leber's optic atrophy.

The term “treatment” refers to both therapeutic treatment andprophylactic or preventative measures, wherein the object is to prevent,retard, or attenuate the related eye disorder as listed above. Those inneed of treatment include those already experiencing the disease orcondition, those prone to having the disease or condition, and those inwhich the disease or condition is to be prevented. The terms “prevent,”“preventing,” and “prevention” refer to delaying or precluding the onsetof a disorder; and/or its attendant symptoms, in a subject or reducing asubject's risk of acquiring a disorder.

The term “subject” refers to an animal, preferably a mammal andincluding primates (e.g., human, monkey, chimpanzee, gorilla, and thelike), The terms “subject” and “patient” are used interchangeably hereinin reference, for example, to a mammalian subject, such as a humanpatient.

Although intravitreal injection is the preferred method ofadministration, other modes of administration are contemplated,including topical, subconjunctival and subtenon administration.

Optic nerve atrophy can be congenital or acquired. If congenital, it isusually hereditary with an onset of deterioration in childhood oradolescence (e.g. Leber's hereditary optic neuropathy (LHON), or Leber'soptic atrophy, dominant optic atrophy or Kjer's optic neuropathy andnumerous less common genetically related syndromes). Alternatively,congenital optic atrophy can be caused by a lack of oxygen duringpregnancy, labor or in the immediate postnatal period. Some drugs takenduring pregnancy are also associated with optic atrophy. Acquired opticatrophies result from multiple etiologies such as decreased blood supplyto the eye or optic nerve (anterior ischemic optic neuropathy orposterior ischemic optic neuropathy), inflammation or swelling withinthe optic nerve (optic neuritis), and pressure against the optic nerve(such as from a tumor or glaucoma). Although less common, it can also berelated to metabolic diseases (e.g., diabetes mellitus), trauma, ortoxicity (caused by methanol, tobacco, or other poisons). It is alsoseen in vitamin B12 deficiency and Paget's disease of bone.

The most common optic neuropathy is glaucomatous optic neuropathy (akaglaucoma), distinguished by a distinctive and progressive excavation ofthe optic nerve head without significant pallor of the remainingneuroretinal rim. In glaucomatous optic neuropathy, retinal ganglioncells (RGCs) die.

The process of RGC death is thought to be biphasic, where a primaryinjury responsible for initiation of damage is followed by a slowersecondary degeneration related to the noxious environment surroundingthe degenerating cells. For example, retinal ischemia as a result ofincreased intraocular pressure may establish a cascade of changes thatultimately results in cell death. Hypoxia leads to excitotoxic levels ofglutamate, which cause a rise in intra-cellular calcium, which in turn,leads to neuronal death due to apoptosis or necrosis. (Kaushik et al.,Neuroprotection in glaucoma. J Postgrad Med. 2003, 49(1):90-5)

Increased intraocular pressure (IOP) (above 22 mmHg or 2.9 kPa) is asignificant risk factor for developing glaucoma. However, significantvariability exists with respect to sensitivity of the optic nerve toincreased IOP with some patients developing nerve damage at a relativelylow pressure, while others may have high pressure for years and yetnever develop damage. Furthermore, while reducing IOP helps preventglaucoma in some at-risk individuals (e.g., those with ocularhypertension) and also prevents progression of glaucoma in someindividuals with existing disease, simply reducing IOP is not alwayseffective. Moreover, achieving adequate pressure lowering may bedifficult or associated with adverse effects (AE). On the other hand,neuroprotection is a process that attempts to preserve the cells thatwere spared during the initial insult, but are still vulnerable todamage. Although not yet available, a neuroprotective agent potentiallycould be of great use in arresting the progression of glaucoma.

Hypoxic injury of the optic nerve is not limited to glaucoma. Ischemicoptic neuropathy (ION) is another important subtype of optic nerveatrophy that includes a variety of disorders associated with ischemia ofthe optic nerve. Posterior Ischemic Optic Neuropathy (PION) is a raremedical condition characterized by ischemic damage to the retrobulbarportion of the optic nerve. (Hayreh S S. Posterior ischaemic opticneuropathy: clinical features, pathogenesis, and management. Eye. 2004,18(11):1188-206).

The more common form of ION, Anterior Ischemic Optic Neuropathy (AION)is the result of disturbances in blood flow through the posteriorciliary arteries leading to ischemic injury of optic nerve axons in theoptic nerve head and subsequent loss of retinal ganglion cells. AION canbe distinguished from PION by the fact that AION occurs spontaneouslyand unilaterally in patients with predisposing anatomy andcardiovascular risk factors. Furthermore, by definition, ION is termedanterior if disc edema is present acutely. (Biousse V, Newman N.J.Neuro-Ophthalmology Illustrated. New York, N.Y.: Thieme MedicalPublishers; 2009).

After glaucoma, AION is the second most common optic nerve-related causeof permanent visual loss in adults. Clinically, AION is of two types:

1. Arteritic AION (A-AION): causes a severe loss of vision and is theprimary cause of vision loss in patients with temporal arteritis (alsocalled giant cell arteritis), a systemic disorder affecting primarilythe elderly characterized by granulomatous (giant cells) inflammation oflarge- and medium-sized arteries. A-AION represents less than 6% of allcases of AION. (Miller et al., Walsh and Hoyt's ClinicalNeuro-Ophthalmology: The Essentials. 2 ed. Philadelphia, Pa.: LippincottWilliams &Wilkins; 2007).

2. Non-Arteritic AION (NAION): includes all other cases of AIONcoincident with cardiovascular risk factors in a patient with “crowded”(i.e., having low cup-to-disc ratio) optic discs. NAION is the mostcommon cause of sudden optic nerve-related vision loss and isresponsible for 95% of all cases of AION.

Mechanistically NAION cases can be broadly classified into two groups:

Thrombosis or embolism of the posterior ciliary arteries or theirsubdivisions—these are rare events in NAION;

Transient poor circulation or no circulation in the blood vessels of theoptic nerve head is the most common cause of NAION (Hayreh S S. Ischemicoptic neuropathy. Prog Retin Eye Res. 2009, 28(1):34-62). Transientlypoor circulation or loss of circulation in the optic nerve head canoccur due to a transient drop in blood pressure which, in susceptiblepersons, can result in NAION. In this etiology of AION, there is noactual blockage of the posterior ciliary arteries. A drop in local bloodpressure in the capillaries of the optic nerve head also may be causedeither by blockage or severe narrowing of the internal carotid arteryand/or the ophthalmic artery, or by a rise in intraocular pressure, orsome combination of these factors. The severity of damage to the opticnerve head depends on the extent and duration of the resulting ischemia,but is usually less extensive and less severe than damage caused bythrombosis or embolism.

NAION typically presents as an abrupt, painless mono-ocular vision loss,though a few patients do experience some discomfort. Visual loss varieswidely, ranging from minor loss of visual acuity to complete blindness.The deterioration of vision is usually discovered upon waking in themorning. (Hayreh et al., Nonarteritic anterior ischemic opticneuropathy: time of onset of visual loss. Am J Ophthalmol. 1997,124(5):641-7).

Visual field defects in NAION are characteristic with patients typicallycomplaining of loss of vision towards the nose and, less commonly,altitudinal loss. Later on, photophobia is a common complaint,particularly in the rare bilateral cases. The optic nerve head acutelyappears edematous, which confirms the anterior nature of this disorder.Hemorrhage on the disc is commonly present.

The estimated mean annual incidence of NAION among persons 50 or olderranged from 2.30 to 10.2 per 100,000, (Johnson L N, Arnold A C.Incidence of nonarteritic and arteritic anterior ischemic opticneuropathy. Population-based study in the state of Missouri and LosAngeles County, California. J Neuroophthalmol. 1994, 14(1):38-44) andmany sources quote the incidence of NAION as about 8,000/year in the US(Hattenhauer et al., Incidence of nonarteritic anterior ischemic opticneuropathy. Am J Ophthalmol. 1997, 123(1):103-7). NAION is primarily adisease of the middle-aged and elderly, although persons at all ages areat risk. It is more prevalent in men and in the white populationrelative to other racial groups. Currently there are no approvedtherapies for NAION.

The neuronal degeneration process can be described in three steps:primary axon damage, concomitant retrograde death of the associatedneuronal cell bodies, and subsequent damage/death of adjacent neurons ina process called “secondary degeneration”. This secondary degenerationoccurs in neurons that initially were not damaged but become exposed tocytokines released during the death of adjacent neurons that experiencedprimary axonal damage. Both primary retrograde cell death followingaxonal damage and secondary degeneration are believed to be mediatedprimarily by apoptosis, or programmed cell death. Accordingly,therapeutic intervention in the form of inhibiting apoptosis could beeffective in protecting neurons following primary axonal damage, andthose that are lost as a result of secondary degeneration. This strategymay be useful even when the initial cause of the disease is not knownbecause it aims at limiting or preventing neuronal damage/death byblocking the underlying cellular mechanism (apoptosis) that gives riseto optic nerve atrophy. (Brao-Osuna et al., New therapeutic systems ofneuroprotectors agents in the treatment of glaucoma. Arch Soc EspOftalmol. 2007, 82(4):191-3).

Direct damage to the ganglion cell body that occurs, for example, duringocclusion of the central artery of the retina, leads to rapid death ofthe nerve in as short as twenty minutes, which is why is it is such acatastrophic disease. In contrast, the site of injury in axogenicdiseases like NAION is the axon, where the process of retrograde celldeath is much slower.

Optic nerve atrophy is defined as the loss of the fibers of the opticnerve, and results from the death of retinal ganglion cells (RGCs).Since they are unable to divide, loss of RGCs results in irreversibleloss of vision. Therefore, therapeutic intervention in diseases thatlead to optic nerve atrophy with neuroprotective agents could preserveRGCs and thereby preserve vision.

The majority of the cases of glaucoma are the form known asprimary-open-angle glaucoma POAG, also called chronic open-angleglaucoma. POAG results from a build up of aqueous humor fluid within theanterior chamber of the eye resulting in intraocular pressure (IOP).Elevated IOP, which can be measured by a “tonometry” test, results fromfluid entering the eye and not enough fluid exiting the eye. Normally,fluid enters the eye by seeping out of the blood vessels in the ciliarybody. This fluid eventually makes its way past the crystalline lens,through the pupil (the central opening in the iris), and into theirido-corneal angle, the anatomical angle formed where the iris and thecornea come together. Then the fluid passes through the trabecularmeshwork in the angle and leaves the eye via the canal of Schlemm.

If excess fluid enters the eye, or if the trabecular meshwork “drain”gets clogged up (for instance, with debris or cells) so that not enoughfluid is leaving the eye, the pressure builds up in what is known as“open angle glaucoma.” Open angle glaucoma also can be caused when theposterior portion of the iris adheres to the anterior surface of thelens creating a “pupillary block”, and preventing intraocular fluid frompassing through the pupil into the anterior chamber.

If the angle between the iris and the cornea is too narrow or is evenclosed, then the fluid backs up, causing increased pressure in what isknown as “closed angle glaucoma.”

Acute angle closure glaucoma (AACG) (also called narrow angle glaucoma)is an ocular emergency with acute presentation, need for immediatetreatment, and well-established anatomic pathology. AACG is defined asat least two of the following symptoms: ocular pain, nausea/vomiting anda history of intermittent blurring of vision with halos; and at leastthree of the following signs: IOP greater than 21 mm Hg, conjunctivalinjection, corneal epithelial edema, mid-dilated nonreactive pupil andshallower chamber in the presence of occlusion.

Primary angle closure glaucoma (PACG) is defined as an occludabledrainage angle and features indicating that trabecular obstruction bythe peripheral iris has occurred with the presence of glaucomatous opticneuropathy.

Untreated glaucoma eventually leads to optic atrophy and blindness.

CASP2 Double-Stranded RNA (dsRNA) QPI-1007

The CASP2 dsRNA QPI-1007 or the sodium salt thereof is a double-stranded(19-base pair duplex), chemically modified, synthetic RNA targetingCaspase 2 mRNA, for example, the mRNA coding sequence (SEQ ID NO:3-5)for human CASP2, and has the following double-stranded structure:

(sense strand; SEQ ID NO: 1) 5′ iB - GCCAGAAUGUGGAACUCCU 3′(antisense strand; SEQ ID NO: 2) 3′ CGGUCUUACACCUUGAGGA 5′wherein each A, C, U, and G is a nucleotide and each consecutivenucleotide is joined to the next nucleotide by a phosphodiester bond;wherein the sense strand comprises, counting from the 5′ terminus, anunmodified ribonucleotide at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17 and 19, a L-deoxycytidine at position 18, andan inverted deoxyabasic 5′ cap (iB); andwherein the antisense strand comprises, counting from the 5′ terminus, a2′-O-Methyl sugar modified ribonucleotide at positions 2, 4, 6, 8, 11,13, 15, 17 and 19 and an unmodified ribonucleotide at positions 1, 3, 5,7, 9, 10, 12, 14, 16 and 18.

The 5′ end of the sense strand of QPI-1007 includes an inverted abasicdeoxyribose sugar that, in addition to conferring resistance to nucleasedegradation, also blocks the ability of the 5′ end of the sense strandto be phosphorylated. Without wishing to be bound to theory, thismodification prevents RNAi-mediated activity of the sense strand. Inaddition, an L-DNA moiety was incorporated at position 18 of the sensestrand to introduce thermodynamic instability in the duplex to favorloading of the antisense strand into RISC. Without wishing to be boundto theory, only the antisense strand of QPI-1007 is capable of elicitingRNAi activity. The antisense strand of QPI-1007 contains multiple 2′-OMethyl (2′-OMe) sugar modified nucleosides at positions 2, 4, 6, 8, 11,13, 15, 17 and 19 that confer nuclease resistance to the antisensestrand and that attenuates potential seed region-mediated off-targetactivity, thereby improving the specificity of QPI-1007 to its intendedtarget, caspasc 2 mRNA (Jackson et al., Position specific chemicalmodification of siRNAs reduces “off-target” transcript silencing. RNA.2006, 12(7):1197-205).

Without wishing to be bound to theory, these chemical modificationsconfer nuclease resistance and mitigate potential off-target activitythat might arise from unwanted RNAi activity elicited by either thesense strand or the seed region of the antisense strand.

Function of Caspase 2

The caspascs are a family of cysteine protcases that play a major rolein apoptosis, or programmed cell death. Twelve human caspases have beenidentified and all initially exist in an inactive (zymogen) state,called procaspases (Logue and Martin, Caspase activation cascades inapoptosis. Biochem Soc Trans. 2008, 36(Pt 1):1-9). External stress ordeath signals, or significant intracellular damage, can triggerapoptosis and subsequent activation of the caspase pathways (Fan et al.,Caspase family proteases and apoptosis. Acta Biochim Biophys Sin(Shanghai). 2005, 37(11):719-27). There are two primary classes ofcaspases in the apoptotic cascade, initiators/activators andexecutioners. Initiators cleave executioner caspases to their activeforms or cleave other proteins in the apoptotic cascade, whichsubsequently results in executioner caspase activation. The activatedexecutioners continue the apoptotic cascade (Logue and Martin, 2008, op.cit; Kumar S. Caspase function in programmed cell death. Cell DeathDiffer. 2007, 14(1):32-43). Caspase 2 was one of the first caspasesidentified, but its role and place in the apoptotic cascade is stilldebated (Logue, 2008, ibid.; Troy C M, Shelanski M L. Caspase-2 redox.Cell Death Differ. 2003, 10(1):101-7). Structurally, caspase 2 can beclassified as an initiator although it has been suggested to have bothinitiator and executioner activity in neurons (Fan et al., 2005, ibid;Kumar S. Caspase function in programmed cell death. Cell Death Differ.2007, 14(1):32-43; Troy and Shelanski, 2003, ibid).

Activation of Caspase 2 in Retinal Ganglion Cells

In NAION, RGC damage begins with the axon rather than the cell body(Levin, 2007 op. cit), and cell death can be prolonged (i.e., overseveral days (Slater et al., Rodent anterior ischemic optic neuropathy(rAION) induces regional retinal ganglion cell apoptosis with a uniquetemporal pattern. Invest Ophthalmol Vis Sci. 2008, 49(8):3671-6). RGCdeath in an ischemic event such as NAION is primarily through apoptosis(Katai N, Yoshimura N. Apoptotic retinal neuronal death byischemia-reperfusion is executed by two distinct caspase familyproteases. Invest Ophthalmol Vis Sci. 1999, 40(11):2697-705; Lam et al.,Apoptosis and caspases after ischemia-reperfusion injury in rat retina.Invest Ophthalmol Vis Sci. 1999, 40(5):967-75; Singh et al.,Cell-specific caspase expression by different neuronal phenotypes intransient retinal ischemia. J Neurochem. 2001, 77(2):466-75). Thecaspases play a major role in apoptosis, and Caspase 2 was shown to beactivated specifically in RGCs in rat models of retinal ischemic insult(Singh et al., 2001, ibid; Kurokawa et al., BDNF diminishes caspase-2but not c-Jun immunoreactivity of neurons in retinal ganglion cell layerafter transient ischemia. Invest Ophthalmol Vis Sci. 1999,40(12):3006-11). In the rat model of transient global retinal ischemia,IVT administration of a pan-caspase inhibitor (Lam et al., 1999, ibid)or a Caspase 2-specific inhibitor (Singh et al., 2001, ibid) resulted inattenuated retinal damage.

RNA Interference Pathway

RNAi is a ubiquitous pathway present in plants and animals and isthought to have evolved as a defense against viral infection. Often,viruses generate long double-stranded RNAs during replication. Longdouble-stranded RNAs are not a natural component of eukaryotic cells. Ariboendonuclease called “DICER” recognizes and cleaves longdouble-stranded RNA into discrete 19-21 base pair double-stranded RNAproducts called small interfering RNAs (siRNAs). An enzyme complexcalled the RNA-induced silencing complex (RISC) utilizes the fragmentsgenerated by DICER as guide sequences to seek out and cleave RNAsmatching the loaded siRNA, thereby distinguishing exogenous viral RNAsfrom self RNA.

Synthetic siRNAs

Manipulation of the RNAi pathway to inhibit expression of endogenousgenes was first demonstrated by Fire and Mello in C. elegans using longdouble-stranded RNA triggers that matched endogenous genes (Fire et al.,Potent and specific genetic interference by double-stranded RNA inCaenorhabditis elegans. Nature. 1998, 391(6669):806-11). Such longdouble-stranded RNAs were processed in these forms by DICER into siRNAs,leading to silencing of the targeted genes. However, use of thispowerful technique in mammals was hampered because long double-strandedRNAs induce a potent innate immune response leading to induction of theinterferon pathway (Robbins et al., siRNA and innate immunity.Oligonucl. 2009, 19(2):89-102).

Elbashir et al., (RNA interference is mediated by 21- and 22-nucleotideRNAs. Genes Dev. 2001, 15(2):188-200) produced 21-nucleotide syntheticsiRNA duplexes matching endogenous mammalian genes and demonstrated thatsuch synthetic siRNAs efficiently loaded into RISC (thereby bypassingDICER processing) leading to RNAi-mediated gene silencing in humancells, and that these small synthetic oligonucleotides did not initiatea strong interferon response as seen previously with longerdouble-stranded RNAs. However, more recent work has shown that somesynthetic siRNAs can activate components of the innate immune system,specifically Toll-like Receptor 3 (Kleinman, et al., Sequence- andtarget-independent angiogenesis suppression by siRNA via TLR3. Nature.2008, 452(7187):591-7). Chemical modification of one or both of the RNAstrands can attenuate, or even abrogate, immune system activation.

Pharmacology of QPI-1007

The pharmacology program for QPI-1007 included in vitro studies whichdemonstrated RNAi-mediated reduction of Caspase 2 mRNA, cleavage ofCaspase 2 mRNa (shown by RACE) and reduced potential for off-targetactivity; and animal studies which demonstrated i) uptake offluorescent-labeled siRNA in RGCs following intravitreal (IVT)administration, ii) RNAi-mediated mechanism of action of QPI-1007 inocular tissues harvested after IVT administration, iii) efficacy ofQPI-1007 in three animal models: a rat ocular hypertension model ofglaucoma, and two models of retrograde RGC cell death that arecharacteristic of RGC death in NAION, the optic nerve axotomy and opticnerve crush models.

RNAi activity of QPI-1007 was assessed in vitro following transfectionof QPI-1007 siRNA at various concentrations into human (HeLa) and rat(PC12) cells. Dose-dependent reduction of Caspase 2 mRNA levels wasobserved in both cell types. The potential of QPI-1007 to inhibitunintended targets was evaluated in a cell culture system. Datademonstrate that the potential for QPI-1007 to elicit substantialoff-target effects is low.

In the rat axotomy and optic nerve crush (ONC) models of retrograde RGCcell death, IVT administration of siRNA resulted in significantprotection of RGCs. In the rat axotomy model, two 10 μg/eye (10microgram/eye) IVT administrations of CASP2 siRNA more than doubled theRGC survival rate two weeks post-axotomy. IVT administration of QPI-1007in the rat ONC model led to dose-dependent protection of RGCs, withcomplete preservation of RGCs 7 days post-injury at doses of 20 and 35μg/eye. In the rat intraocular hypertension model of glaucoma, a single20 μg/eye IVT administration of QPI-1007 two weeks following inductionof increased intraocular pressure afforded significant protection ofRGCs.

Overall, the nonclinical pharmacology studies provide evidence of thepharmacological activity of QPI-1007 in a variety of in vitro and invivo systems. Taken together, these data support the clinicaldevelopment of QPI-1007 as a neuroprotectant for the treatment of oculardiseases, disorders and injury, such as nonarteritic anterior ischemicoptic neuropathy and other optic neuropathies that result in the deathof retinal ganglion cells.

Pharmaceutical Compositions, Kits, and Containers

Provided herein are compositions, kits, containers and formulations thatinclude the double-stranded RNA compound provided herein foradministering to a patient, preferably to a patient's eye.

In certain embodiments, the composition is administered in combinationwith an anesthetic. In some embodiments, the anesthetic is a topicalanesthetic suitable for administration to the human eye.

In certain embodiments, the composition is administered in combinationwith a broad-spectrum microbiocide. In some embodiments thebroad-spectrum microbiocide is a broad-spectrum topical microbiocide. Invarious embodiments, the broad-spectrum topical microbiocide.

In certain embodiments, a topical anesthetic and a broad-spectrumtopical microbiocide are applied to the eye prior to administration ofthe composition.

In certain embodiments, the composition is administered in combinationwith an antibiotic. In various embodiments, the antibiotic is anantibiotic suitable for administration to the eye. In variousembodiments, the antibiotic is formulated as eye drops. In certainembodiments, the antibiotic is administered to the patient following theadministration of the composition.

In certain embodiment, the composition further comprises abroad-spectrum microbiocide, an antibiotic or a combination thereof.

As provided herein, a kit may include at least one container and atleast one label. Suitable containers include, for example, bottles,vials, syringes, and test tubes. The containers can be formed from avariety of materials such as glass, metal or plastic. The container canhold the compound. The container can alternatively hold a compositioncomprising the compound.

A kit may further include a second container that includes apharmaceutically acceptable buffer, such as phosphate-buffered saline,Ringer's solution and/or dextrose solution. It can further include othermaterials desirable from a commercial and user standpoint, includingother buffers, diluents, filters, stirrers, needles, syringes, and/orpackage inserts with indications and/or instructions for use.

The unit dosage ampoules or multidose containers, in which the compoundis packaged prior to use, may include an hermetically sealed containerenclosing an amount of the compound suitable for a pharmaceuticallyeffective dose thereof, or multiples of an effective dose. The compoundis packaged as a sterile formulation, and the hermetically sealedcontainer is designed to preserve sterility of the formulation untiluse.

The container which includes the compound may further include a packagethat is labeled, and the label may bear a notice in the form prescribedby a governmental agency, for example the Food and Drug Administration,which notice is reflective of approval by the agency under Federal law,of the manufacture, use, or sale of the compound therein for humanadministration.

Federal law requires that the use of pharmaceutical compositions in thetherapy of humans be approved by an agency of the Federal government. Inthe United States, enforcement is the responsibility of the Food andDrug Administration, which issues appropriate regulations for securingsuch approval, detailed in 21 U.S.C. section 301-392. Similar approvalis required by most foreign countries. Regulations vary from country tocountry, but individual procedures are well known to those in the artand the compositions and methods provided herein preferably complyaccordingly.

EXAMPLES

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present invention toits fullest extent. The following preferred specific embodiments are,therefore, to be construed as merely illustrative, and not limitative ofthe claimed invention in any way.

Example 1: Test Compound

The test compound QPI-1007 is a sodium salt of a double-stranded RNAcompound having the structure:

(sense strand; SEQ ID NO: 1) 5′ iB - GCCAGAAUGUGGAACUCCU 3′(antisense strand; SEQ ID NO: 2) 3′ CGGUCUUACACCUUGAGGA 5′wherein each A, C, U, and G is a nucleotide and each consecutivenucleotide is joined to the next nucleotide by a phosphodiester bond;wherein the sense strand comprises, counting from the 5′ terminus, anunmodified ribonucleotide at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17 and 19, a L-deoxycytidine at position 18, andan inverted deoxyabasic 5′ cap; andwherein the antisense strand comprises, counting from the 5′ terminus, a2′-O-Me (2′-O-Methyl, 2′ methoxy) sugar modified ribonucleotide atpositions 2, 4, 6, 8, 11, 13, 15, 17 and 19 and an unmodifiedribonucleotide at positions 1, 3, 5, 7, 9, 10, 12, 14, 16 and 18;

The Molecular Formula of QPI-1007 Sodium Salt is: C(375) H(439) N(143)Na(37) O(266) P(37).

The Molecular Weight of QPI-1007 Sodium Salt is: 13,202 Da.

Example 2: First in Human Open Label Phase I Clinical Trial of QPI-1007in NAION Patients

The study was designed to enroll up to 66 patients in two separatestrata as described below.

Dosing of the first cohort of Stratum I commenced at the first doselevel of 0.2 mg in a single eye and subsequent Stratum I cohorts wererecruited sequentially to receive escalating doses of QPI-1007 (FIG. 1).In this stratum, no more than one patient received QPI-1007 in any24-hour period.

Following administration of QPI-1007 to the last patient in a cohort inStratum I, a 7-day follow-up period and a review of available safetydata is required before the first patient in the next cohort receivesQP1-1007.

FIG. 2 provides a flow chart outlining the Dose Escalation program.

Inclusion Criteria: Patients meet the following inclusion criteria:

Stratum I

1. “Legally blind” in the study eye using the US definition of visualacuity of less than or equal to 20/200 or visual field restricted toless than 20 degrees. This level of visual function must have beenstable for a minimum of 6 months prior to screening. Stable is definedas the same visual acuity score at screening and at least 6 monthspreviously using the same visual acuity test (ETDRS or Snellen).

2. Blindness is the result of an irreversible condition affecting theposterior segment of the study eye. These conditions include, but arenot limited to: Retinal degeneration, Optic neuritis End stage glaucoma,Optic nerve atrophy due to any etiology, Leber's hereditary opticneuropathy with onset at least 2 years prior to screening.

3. Clear ocular media and able to undergo adequate pupil dilation toallow a good fundus examination.

4. Both visual acuity and visual field in the non-study eye are betterthan or equal to the study eye at screening.

Stratum II

1. Positive diagnosis of NAION with symptom onset within 14 days priorto planned dosing with QPI-1007.

The NAION diagnosis required all of the following: Disc edema, Visualfield defects in the study eye consistent with optic neuropathy and meandeviation on Humphrey SITA standard 24-2 worse than −3.0 dB, Relativeafferent pupillary defect (if the study eye is the first eye affected).

2. Best-corrected visual acuity in the study eye is worse than or equalto 20/64 and better than or equal to light perception.

3. Best corrected visual acuity and visual field in the non-study eyeare better than or equal to the study eye at screening.

Test Product, Dose, and Mode of Administration:

Test Product: The active pharmaceutical ingredient of QPI-1007 is asodium salt of a double-stranded (19-base pair), chemically modified,synthetic siRNA targeting Caspasc 2 mRNA. QPI-1007 has been manufacturedin accordance with Good Manufacturing Practices (GMP).

The drug product, “QPI-1007 Injection”, is a preservative-free, sterilesolution formulated at 60 mg/mL in phosphate-buffered saline. QPI-1007was supplied as a sterile solution for IVT injection in a 2 mL Type Iglass vials sealed with Teflon-coated butyl rubber stoppers withaluminum flip-off overseals. Each vial was intended for single use andcontains volume sufficient to dose concentrations defined by theprotocol. QPI-1007 was diluted to the target concentration with anappropriate volume of diluent. The final concentration to be preparedwas determined by the intended dose level.

QPI-1007 Injection was stored refrigerated at 2-8° C., protected fromlight. The solution was warmed to room temperature prior to use.

Mode of Administration

In the study, QPI-1007 was administered via intravitreal (IVT)injection. QPI-1007 was diluted with a sterile saline solution suitablefor injection into the eye to achieve the correct concentration to bedelivered by IVT injection.

Preparation for administration was done using aseptic techniques. TheIVT injection procedure was carried out under controlled asepticconditions, which included the use of sterile gloves, a sterile drape,and a sterile eyelid speculum (or equivalent). Adequate topicalanesthetic and a broad-spectrum topical microbiocide were given prior tothe injection.

Following the IVT injection, patients were monitored for elevation ofintraocular pressure, decreased optic nerve head perfusion and forpossible injection complications (vitreous hemorrhage, retinal tears).Additionally, patients had to report any symptoms suggestive ofendophthalmitis, such as ocular pain, swelling, redness, haze andgradual loss of vision immediately.

Patients were prescribed antibiotic eye drops following administrationof QPI-1007.

Detailed guidelines on IVT injection procedures are known in the art(Aiello, et al., Evolving guidelines for intravitreous injections.Retina. 2004, 24(5 Suppl):S3-19; Brucker (Brucker A J. Maximizing theSafety of Intravitreal Injections. Medscape Ophthalmology [Internet].2006 7(2)).

Dosing

Stratum I: QPI-1007 was administered as a single IVT injection topatients in each cohort according to the following schedule in Table 1.

TABLE 1 Cohort N patients Dose per Injection (mg) 1 3 to 6 0.2 2 3 to 6Up to 0.6 3 3 to 6 Up to 1.2 4 3 to 6 Up to 2.4 5 3 to 6 Up to 4.8 6 3to 6 Up to 6.0

Stratum II: QPI-1007 was administered as a single IVT injection topatients in each cohort according to the following schedule in Table 2.

TABLE 2 Dose per Cohort N Patients Injection (mg) 1 (following StratumI, Cohort 3) Up to 10 Up to 1.2 2 (following Stratum I, Cohort 4) Up to10 Up to 2.4 3 (following Stratum I & II, Cohorts 1 & 2) Up to 10 >1.2and ≤6.0Duration of Treatment and Follow-Up:

Stratum I

In Stratum I all screening procedures were performed within 28 daysprior to IVT injection of QPI-1007 (on Study Day 0).

At screening, patients underwent an ophthalmic evaluation (both eyes)including best corrected visual acuity (BCVA) assessment, visual fieldassessment, tonometry, slit lamp examination of the anterior segment andslit lamp/ophthalmoscope examination of the posterior segment, fundusphotography (FP), and optical coherence tomography (OCT) of the maculaand of the peripapillary retinal nerve fiber layer.

Prior to IVT injection of QPI-1007 on Study Day 0, the patient underwenta BCVA assessment and tonometry in both eyes. 30 minutes following IVTinjection the patient's study eye was examined by tonometry to determineintraocular pressure, and by slit-lamp/ophthalmoscope to determine thestatus of optic nerve perfusion and to check for any retinal hemorrhageor tears. Patients were monitored in the clinic for 4 hours followinginjection as per the schedule of events. Patients returned to the clinicthe day following IVT injection (Study Day 1) for a safety assessment ofboth eyes including BCVA assessment, tonometry, slit lamp examination ofthe anterior segment and slit lamp/ophthalmoscope examination of theposterior segment.

On Study Days 7, 14 and 28 and at Study Weeks 8 and 12 after IVTinjection of QPI-1007, patients underwent an ophthalmic evaluation (botheyes) including BCVA assessment, tonometry and slit lamp examination ofthe anterior segment and slit lamp/ophthalmoscope examination of theposterior segment. In addition, patients underwent a visual fieldassessment, FP and OCT of the macula and of the peripapillary retinalnerve fiber layer at Study Week 12 only.

Follow-up visits occur at Study Months 6 and 12 after IVT injection ofQPI-1007. Patients underwent an ophthalmic evaluation (both eyes)including BCVA assessment, tonometry and slit lamp examination of theanterior segment and slit lamp/ophthalmoscope examination of theposterior segment.

Patient safety was monitored during the study by evaluation of vitalsigns, clinical laboratory testing, physical examinations, collection ofadverse events (AEs) and review of concomitant medications.

Stratum II

In Stratum II, all screening procedures were performed within 48 hoursprior to IVT injection of QPI-1007 (on Study Day 0), and were conductedon the same day as the IVT injection. Patients received the QPI-1007injection no more than 14 days from the onset of NAION symptoms.

At screening, patients underwent an ophthalmic evaluation (both eyes)including BCVA assessment, visual field assessment, tonometry, slit lampexamination of the anterior segment and slit lamp/ophthalmoscopeexamination of the posterior segment, FP and OCT of the macula and ofthe peripapillary retinal nerve fiber layer.

Prior to IVT injection of QPI-1007 on Study Day 0, the patient underwenttonometry (both eyes). 30 minutes following IVT injection, the patient'sstudy eye was examined by tonometry to determine intraocular pressureand by slit-lamp/ophthalmoscope to determine the status of optic nerveperfusion and to check for any retinal hemorrhage or tears. Patientswere monitored in the clinic for 4 hours following IVT injection as perthe schedule of events. Patients returned to the clinic the next dayfollowing IVT injection (Study Day 1) for a safety assessment of botheyes including BCVA assessment, tonometry, slit lamp examination of theanterior segment and slit lamp/ophthalmoscope examination of theposterior segment.

On Study Days 7, 14 and 28 and at Study Weeks 8 and 12 after IVTinjection of QPI-1007, patients underwent an ophthalmic evaluation (botheyes) including BCVA assessment, tonometry, slit lamp examination of theanterior segment and slit lamp/ophthalmoscope examination of theposterior segment, FP and OCT of the macula and of the peripapillaryretinal nerve fiber layer. In addition, on Study Days 7 and 28 and atStudy Week 12, patients underwent a visual field assessment.

Follow-up visits occur at Study Months 6 and 12 after IVT injection ofQPI-1007. Patients underwent an ophthalmic evaluation (both eyes)including BCVA assessment, tonometry, slit lamp examination of theanterior segment and slit lamp/ophthalmoscope examination of theposterior segment, FP and OCT of the macula and of the peripapillaryretinal nerve fiber layer. In addition, at Study Month 6 only, patientsreceived a visual field assessment.

Patient safety was monitored on an ongoing basis during the studyincluding but not limited to study assessments including evaluation ofvital signs, clinical laboratory testing, physical examinations,collection of AEs and review of concomitant medications.

Ophthalmic Evaluations

The following ophthalmic evaluations were performed for both eyes forpatients enrolled in Stratum 1:

Best Corrected Visual Acuity (BCVA) was assessed using the ETDRS chartstarting at 4 meters at Screening and on Study Days 0 (pre-injection, ifScreening assessment was conducted more than 48 hours previously), 1, 7,14 and 28, Study Weeks 8 and 12/ET and Study Months 6 and 12.

Visual field was assessed using the SITA Standard algorithm at Screeningand Study Week 12/ET.

Slit lamp examination of the anterior segment of the eye was performedwithout pupillary dilation, whenever possible. Any abnormalities of theanterior chamber, eyelids, conjunctivae, iris, lens and cornea weredocumented at Screening and on Study Days 1, 7, 14 and 28, Study Weeks 8and 12/ET and Study Months 6 and 12. Any anterior chamber inflammation,phakic status and posterior lens capsule status were noted.

Intraocular pressure (IOP) was measured using Goldmann applanationtonometry at Screening and on Study Days 0 (pre-injection), 1, 7, 14 and28, Study Weeks 8 and 12/ET and Study Months 6 and 12. Wheneverpossible, tonometry was performed prior to pupillary dilation.

Slit lamp/ophthalmoscope examination of the posterior segment of the eyewas performed after pupillary dilation to examine the vitreous body,optic nerve head, macular and peripheral retina and fundus at Screeningand on Study Days 1, 7, 14 and 28, Study Weeks 8 and 12/ET and StudyMonths 6 and 12. Any vitreous inflammation was noted.

Fundus photographs were obtained at Screening and Study Week 12/ET only.

Optical coherence tomography of the macula and of the peripapillaryretinal nerve fiber layer was obtained at Screening and Study Week 12/ETonly.

In addition, the following ophthalmic evaluations were performed in thestudy eye only 30 minutes post-injection on Study Day 0 for patientsenrolled in Stratum I: Slit lamp examination of the anterior segment,Slit lamp/ophthalmoscope examination of the posterior segment, IOPmeasured by Goldmann applanation tonometry.

The following ophthalmic evaluations were performed for both eyes forpatients enrolled in Stratum II:

Best Corrected Visual Acuity (BCVA) was assessed using the ETDRS chartstarting at 4 meters at Screening and on Study Days 1, 7, 14 and 28,Study Weeks 8 and 12/ET and Study Months 6 and 12.

Visual field was assessed at Screening, Study Days 7 and 28, Study Week12/ET, and Study Month 6.

Slit lamp examination of the anterior segment of the eye was performedwithout pupillary dilation, whenever possible. Any abnormalities of theanterior chamber, eyelids, conjunctivae, iris, lens and cornea weredocumented at Screening and on Study Days 1, 7, 14 and 28, Study Weeks 8and 12/ET and Study Months 6 and 12. Any anterior chamber inflammation,phakic status and posterior lens capsule status were noted.

Intraocular pressure (TOP) was measured using Goldmann applanationtonometry at Screening and on Study Days 0 (pre-injection), 1, 7, 14 and28, Study Weeks 8 and 12/ET and Study Months 6 and 12. Wheneverpossible, tonometry was performed prior to pupillary dilation.

Slit lamp/ophthalmoscope examination of the posterior segment of the eyewas performed after pupillary dilation to examine the vitreous body,optic nerve head, macular and peripheral retina and fundus at Screeningand on Study Days 1, 7, 14 and 28, Study Weeks 8 and 12/ET and StudyMonths 6 and 12. Any vitreous inflammation was noted.

Fundus photographs were obtained at Screening and on Study Days 7, 14,and 28, Study Weeks 8 and 12/ET, and are obtained at Study Months 6 and12.

Optical coherence tomography of the macula and of the peripapillaryretinal nerve fiber layer was obtained at Screening and on Study Days 7,14, and 28, Study Weeks 8 and 12/ET, and are obtained at Study Months 6and 12.

In addition, the following ophthalmic evaluations were performed in thestudy eye only 30 minutes post-injection on Study Day 0 for patientsenrolled in Stratum II: Slit lamp examination of the anterior segment,Slit lamp/ophthalmoscope examination of the posterior segment, TOPmeasured using Goldmann applanation tonometry.

TABLE 3 QPI-1007 Study Schedule - Stratum I - Optic Nerve AtrophyPatients Active Study Period Screening Study Visits Follow Follow (≤28days of Day Day Day Day Day Week Week up month up month ProceduresInjection) 0 1 7 14 28 8 12/ET 6 12 Written Informed X ConsentDemographics X Physical Exam X X Medical History X Medication X HistoryClinical Labs X X Pregnancy Test X X¹ X Assess Eligibility X X  CriteriaAdminister X  QPI-1007 Vital Signs X X² X X X X X X BCVA X X¹ X X X X XX X X Visual Field X X Slit lamp exam X X³ X X X X X X X X (anter.segment) IOP/Goldmann X X⁴ X X X X X X X X Applanation Tonometry Slitlamp/ X X³ X X X X X X X X ophthalmoscope exam (post segment) Fundus X Xphotography OCT X X PK Blood draw⁷ X⁵  X⁶ X X Concomitant X  X X X X X XMedications All AEs X  X X X X X X Ocular AEs of X X special interestonly ¹Pre-injection only - do not repeat if screening was conductedwithin 48 hours of QPI-1007 injection ²Pre-injection, 30 minutes and 4hours post-injection ³30 minutes post-injection (study eye only)⁴Pre-injection (both eyes) and 30 minutes post-injection (study eyeonly) ⁵Pre-injection, 1 and 4 hours post-injection ⁶24 hourspost-injection ⁷For the first 3 patients of every cohort only

TABLE 4 QPI-1007 Study Schedule - Stratum II- NAION Patients ActiveStudy Period Screening¹ Study Visits Follow Follow (≤48 days of Day DayDay Day Day Week Week up month up month Procedures Injection) 0 1 7 1428 8 12/ET 6 12 Written Informed X Consent Demographics X Physical ExamX X Medical History X Medication X History Clinical Labs X X PregnancyTest X X Assess Eligibility X Criteria Administer X QPI-1007 Vital SignsX  X² X X X X X X BCVA X X X X X X X X X Visual Field X X X X X Slitlamp exam X  X³ X X X X X X X X (anter. segment) IOP/Goldmann X  X⁴ X XX X X X X X Applanation Tonometry Slit lamp/ X  X³ X X X X X X X Xophthalmoscope exam (posterior segment) Fundus X X X X X X X Xphotography OCT X X X X X X X X PK Blood Draw⁷  X⁵  X⁶ X X Concomitant XX X X X X X Medications All AEs X X X X X X X Ocular AEs of X X specialinterest only ¹Screening and Study Day 0 may occur on the same day²Pre-injection, 30 minutes and 4 hours post-injection ³30 minutespost-injection (study eye only) ⁴Pre-injection (both eyes) and 30minutes post-injection (study eye only) ⁵Pre-injection, 1 and 4 hourspost-injection ⁶24 hours post-injection ⁷For the first 3 patients ofevery cohort onlyResults:

Subjects with long-standing low vision due to retinal or optic nervepathology and subjects with acute NAION were studied in a 1 year,2-stratum, Phase I, multi-center, open-label, dose escalation study todetermine safety, tolerability and the structural and functional changesafter a single IVT injection of QP1-1007, a synthetic, chemicallymodified siRNA that inhibits expression of caspase 2.

Low-vision subjects with visual acuity (VA)≤20/200 (Stratum I) and NAIONsubjects with ≤20/40 and symptom onset within 28 days prior to the studydrug injection (Stratum II-S2) were enrolled in 6 cohorts (0.2-6 mg) and3 cohorts (1.2, 2.4 and 6 mg), respectively. After receiving a singleIVT injection, subjects were evaluated for VA, visual field (VF) andretinal nerve fiber layer (RNFL) thickness at days 1, 7, 14, 28, andmonths 2, 3, 6, 12.

48 subjects (18 low vision, 30 NAION) were enrolled. All expected studyvisits are complete for all subjects, except the final follow-up visit(Month 12) for the last enrolled cohort (S2, 6 mg). Available data fromboth strata through Month 3 (n=48), Month 6 (n=48) and Month 12 (n=38)were analyzed. 261 of 273 adverse events (AEs) were of mild-to-moderateseverity. There were no serious AEs. The most common AEs wereconjunctival hemorrhage (n=29), conjunctival chemosis (n=11), and eyepain (n=11). Among 28 NAION subjects in S2 with on-chart VA, maximum VAgain was at Month 2 (mean±SD: 16.4±10.4 letters). The proportion ofsubjects in S2 (FIG. 1) improving by ≥3 lines at Months 3 and 6 were53.6% (n=15), and 50.0% (n=14) compared with 39.7% (n=48), and 42.6%(n=52) of Ischemic Optic Neuropathy Decompression TrialIONDT) historicalcontrols (p=0.2 and 0.5, respectively; Fisher exact test) [IschemicOptic Neuropathy Decompression Trial, Arch Ophthalmol. 2000;118(6):793-797]. Of all the S2 subjects with available follow-up data,no subject lost ≥3 lines of VA compared with 9.1% (n=11), 14.8% (n=18),and 15.8% (n=18) at Months 3, 6 and 12, respectively, in the IONDThistorical controls. VF mean defect was comparable to baseline. Decreasein RNFL thickness was similar to historical controls [Contreras et al.,2007].

CONCLUSION

A single IVT injection of QPI-1007 was well tolerated in subjects withlong-standing low vision or acute NAION. Patients treated by a singleIVT injection of QPI-1007 were protected from further loss of visualacuity compared to published historical data on untreated NAION patientswith similar initial disease severity.

Example 3: A Phase II Pivotal Randomized, Double Masked, Sham-ControlledTrial of QPI 1007 Delivered by a Intravitreal Injections to Patientswith Acute Non Arteritic Anterior Ischemic Optic Neuropathy (NAION)

Study Design:

This is a double masked, randomized sham-controlled efficacy and safetystudy. The study will enroll up to 240 patients with an acute NAION.Patients will be randomized into one of 3 groups in a 1:1:1 ratio. Twogroups will receive treatment with QPI-1007, and the third group willreceive a sham injection.

Patients randomized to one of the treatment groups will receive monthlyintravitreal injections of either 2.4 mg or 6.0 mg of QPI-1007 in thestudy eye on the day of randomization and at study month 1, 2, 3, 4 and5. Subjects will be stratified by baseline (Day 0) BCVA score (>20/64and ≤20/64) and by country. Patients randomized to the sham-control armwill receive a sham-injection in the study eye on the day ofrandomization and study month 1, 2, 3, 4 and 5. Patients, techniciansand Investigators will be masked to treatment arm. Only the injectingphysician will be unmasked, but will not be involved in patientevaluations other than the immediate post-injection patient evaluation.

Patients will be followed monthly for 6 months, with a final follow upvisit at month 12. Study procedures are listed in the Schedule of EventsTable below.

Inclusion Criteria:

Patients must meet the following inclusion criteria:

1. Positive diagnosis of Non-Arteritic AION (NAION) with symptom onsetwithin 28 days prior to planned dosing with QPI-1007. The NAIONdiagnosis requires all of the following: Disc edema, Visual fielddefects in the study eye consistent with optic neuropathy and meandeviation on Humphrey SITA standard 24-2 worse than −3.0 dB and Relativeafferent pupillary defect (if the study eye is the first eye affected)

2. Best-corrected visual acuity score in the study eye is worse than orequal to 20/20 and better than or equal to 20/400 Snellen equivalents,measured on the ETDRS chart

3. 50 years old or older at screening.

4. Clear ocular media and able to undergo adequate pupil dilation toallow a good fundus examination.

5. Capable of giving written informed consent.

6. Willing and able to comply with the study procedures and visitschedule, including follow-up visits.

7. Female patients must be: (1) post-menopausal (2) surgically sterile,or (3) using an effective means of contraception which will be continueduntil the Study Month 6 visit with a negative pregnancy test within 48hours prior to administration of QPI-1007. Male patients with femalepartners of child bearing potential must agree to use an effective meansof contraception which will be continued until the Study Month 6 visit.Note: For the purpose of this study, post-menopausal is defined as theabsence of menses for at least one year and a serum FSH level ≥20 IU/L.Investigators can determine if a serum FSH level is required to provepost-menopausal status. A woman is considered to be surgicallysterilized if she has had a bilateral tubal ligation for at least 6months prior to administration of QPI-1007, bilateral oophorectomy, orcomplete hysterectomy. Effective means of contraception include use ofone of the following: hormonal contraceptives (oral, implant,transdermal patch, or injection) at a stable dose for at least 3 monthsprior to administration of QPI-1007, barrier (condom with spermicide,diaphragm with spermicide), IUD, or a male patient/partner who has beenvasectomized for at least 6 months prior to administration of QPI-1007.

Test Product, Dose, and Mode of Administration

QPI-1007 is a double-stranded (19-base pair), chemically-modified,synthetic siRNA targeting caspase 2 mRNA and is designed to temporarilyinhibit the expression of caspase 2. QPI-1007 will be supplied as asterile solution for IVT injection in a 2 mL glass vial. The drugproduct will be diluted to the target concentration with an appropriatevolume of diluent. QPI-1007 will be administered as an IVT injection.All IVT injections will be at the same volume of injection (100 μl).There is no reference therapy administered in this study.

Duration of Treatment and Follow-Up

Patients must receive the first QPI-1007 injection no more than 28 daysfrom the onset of NAION symptoms. If both eyes have NAION and areeligible for the study enrollment, the eye with the worse VA will bechosen as the study eye. If both eyes have the same VA, the eye chosenby the patient and agreed by the Investigator will be assigned as thestudy eye.

At screening, patients will undergo an ophthalmic evaluation (both eyes)including BCVA assessment, visual field assessment, tonometry, slit lampexamination of the anterior segment and slit lamp/ophthalmoscopeexamination of the posterior segment, fundus photo (FP) and SD-OCT ofthe macula and of the peripapillary retinal nerve fiber layer.

Prior to IVT injection of QPI-1007, the patient will undergo BCVAassessment and tonometry (both eyes), and a comprehensive anterior andposterior segment eye exam, as detailed in the events table below.Immediately after the IVT injection, the patient's injected eye will beexamined for optic nerve head perfusion. Within 30 minutes following IVTinjection, the patient's study eye will be examined by tonometry todetermine intraocular pressure and by slit-lamp/ophthalmoscope todetermine the status of optic nerve perfusion and to check for anyretinal hemorrhage or tears. Patients will be monitored in the clinicfor up to 4 hours following IVT injection as per the schedule of events.

On Study Days 7, and Study Months 1, 2, 3, 4, 6 and 12, patients willundergo an ophthalmic evaluation (both eyes) including BCVA assessment,tonometry, slit lamp examination of the anterior segment and slitlamp/ophthalmoscope examination of the posterior segment. In addition,on Study Day 0, and Study Months 3, 6 and 12, patients will undergo FP,color vision testing, contrast sensitivity testing and SD-OCT of themacula and of the peripapillary retinal nerve fiber layer. On Study Day0 and Study Months 6 and 12 patients will undergo a visual fieldassessment.

SD-OCT and visual field assessments will be sent to a central readingcenter for evaluation.

Patient safety will be monitored on an ongoing basis during the studyincluding but not limited to study assessments including evaluation ofvital signs, clinical laboratory testing, physical examinations, andcollection of reported AEs and review of concomitant medications.

TABLE 5 Schedule of Events WEEK/Month Screening Day 0 Day 7 Month 1Month 2 Month 3 Month 4 Month 5 Month 6 Month 12 VISIT 1 2 3 4 5 6 7 8 910 Sign Informed Consent X Demographics X Medical/Ophthalmic History X XPhysical Exam X Vital Signs and Weight X X X X X X X X X X Clinical LabsX X X Pregnancy test X X X X X X X X X EKG X X X BCVA (ETDRS) X X X X XX X X X X RAPD X X X X X X X X X X Color Vision X X X X X Contrastsensitivity X X X X X Visual Field X X X X (Humphrey 24-2 protocol) Slitlamp exam (anterior X X X X X X X X X X segment) IOP X X X X X X X X X XPosterior segment evaluation X X X X X X X X X X Fundus Photography X XX X SD-OCT X X X X IVT 2.4 or 6 or Sham Injection X X X X X XPost-Injection posterior X X X X X X segment evaluation Post-InjectionIOP X X X X X X Concomitant medication X X X X X X X X X X AdverseEvents X X X X X X X X X

Results: According to the results that are obtained in this study,multiple monthly IVT injections of QPI-1007 are well tolerated insubjects with acute NAION. Patients treated by a multiple monthly IVTinjections of QPI-1007 are protected from further loss of visual acuitycompared to published historical data on untreated NAION patients withsimilar initial disease severity.

Although the above examples have illustrated particular ways of carryingout embodiments of the invention, in practice persons skilled in the artwill appreciate alternative ways of carrying out embodiments of theinvention, which are not shown explicitly herein. It should beunderstood that the present disclosure is to be considered as anexemplification of the principles of this invention and is not intendedto limit the invention to the embodiments illustrated.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, equivalents of the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

The invention claimed is:
 1. A method of treating a human patientsuffering from or at risk for developing any one or more of primary openangle glaucoma (POAG), normal-tension glaucoma, primary angle-closureglaucoma (PACG), acute angle-closure glaucoma (AACG), angle-closureglaucoma, primary angle closure (PAC), or acute angle closure (AAC), themethod comprising administering to the patient's eye via intravitreal(IVT) injection a double-stranded ribonucleic acid (dsRNA) compound thatdown-regulates CASP2 expression, wherein the double-stranded RNAcompound has the structure: (sense strand; SEQ ID NO: 1) 5′iB - GCCAGAAUGUGGAACUCCU 3′ (antisense strand; SEQ ID NO: 2) 3′CGGUCUUACACCUUGAGGA 5′

wherein each A, C, U, and G is a nucleotide and each consecutivenucleotide is joined to the next nucleotide by a phosphodiester bond;wherein each nucleotide is independently an unmodified ribonucleotide, a2′-O-Methyl sugar modified ribonucleotide, or an L-DNA nucleotide;wherein the sense strand comprises, counting from the 5′ terminus, anunmodified ribonucleotide at each of positions 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, and 19 and a L-deoxycytidine atposition 18, and an inverted abasic deoxyribose cap (iB) covalentlybound to the 5′ terminus; wherein the antisense strand comprises,counting from the 5′ terminus, a 2′-O-Methyl sugar modifiedribonucleotide at each of positions 2, 4, 6, 8, 11, 13, 15, 17, and 19and an unmodified ribonucleotide at each of positions 1, 3, 5, 7, 9, 10,12, 14, 16, and 18; or a pharmaceutically acceptable salt thereof; andwherein the compound is administered to the patient's eye at a dose ofabout 1.2 mg to about 3.0 mg per eye.
 2. The method of claim 1, whereinthe treatment comprises multiple administrations of the compound.
 3. Themethod of claim 2, wherein the IVT injections are administered sixtimes.
 4. The method of claim 2, wherein the multiple administrationsoccur at regular intervals.
 5. The method of claim 2, wherein thecompound is administered as six IVT injections at intervals about onemonth apart.
 6. The method of claim 1, wherein the pharmaceuticallyacceptable salt is a sodium salt.
 7. The method of claim 1, wherein thecompound is administered to the patient's eye as a liquid compositioncomprising a pharmaceutically acceptable carrier.
 8. The methodaccording to claim 7, wherein a volume of a single dose IVT injection isbetween about 20 μl to about 200 μl.
 9. The method according to claim 7,wherein a volume of a single dose IVT injection is between about 50 μlto about 100 μl.